Sat, 26 Jul 2008 07:44:15 BST CiteULike: neils's Knapp http://www.citeulike.org/user/neils/author/Knapp CiteULike.org en-gb Copyright © 2004-2008 citeulike.org http://www.citeulike.org/user/neils/article/2321043 <i>The EMBO Journal, Vol. aop, No. current. (31 January 2008)</i> Activation segment dimerization: a mechanism for kinase autophosphorylation of non-consensus sites Ashley Pike Peter Rellos Frank Niesen Andrew Turnbull Antony Oliver Sirlester Parker Benjamin Turk Laurence Pearl Stefan Knapp doi:10.1038/emboj.2008.8 The EMBO Journal, Vol. aop, No. current. (31 January 2008) 2008-02-02T00:11:33-00:00 2008 The EMBO Journal 0261-4189 aop current Nature Publishing Group activation dimerization kinase phosphorylation substrate http://www.citeulike.org/user/neils/article/2319613 <i>Nat Methods, Vol. 5, No. 2. (February 2008), pp. 135-146.</i><br /><br />In selecting a method to produce a recombinant protein, a researcher is faced with a bewildering array of choices as to where to start. To facilitate decision-making, we describe a consensus 'what to try first' strategy based on our collective analysis of the expression and purification of over 10,000 different proteins. This review presents methods that could be applied at the outset of any project, a prioritized list of alternate strategies and a list of pitfalls that trip many new investigators. Protein production and purification. S Gräslund P Nordlund J Weigelt J Bray O Gileadi S Knapp U Oppermann C Arrowsmith R Hui J Ming S Dhe-Paganon HW Park A Savchenko A Yee A Edwards R Vincentelli C Cambillau R Kim SH Kim Z Rao Y Shi TC Terwilliger CY Kim LW Hung GS Waldo Y Peleg S Albeck T Unger O Dym J Prilusky JL Sussman RC Stevens SA Lesley IA Wilson A Joachimiak F Collart I Dementieva MI Donnelly WH Eschenfeldt Y Kim L Stols R Wu M Zhou SK Burley JS Emtage JM Sauder D Thompson K Bain J Luz T Gheyi F Zhang S Atwell SC Almo JB Bonanno A Fiser S Swaminathan FW Studier MR Chance A Sali TB Acton R Xiao L Zhao LC Ma JF Hunt L Tong K Cunningham M Inouye S Anderson H Janjua R Shastry CK Ho D Wang H Wang M Jiang GT Montelione DI Stuart RJ Owens S Daenke A Schütz U Heinemann S Yokoyama K Büssow KC Gunsalus doi:10.1038/nmeth.f.202 Nat Methods, Vol. 5, No. 2. (February 2008), pp. 135-146. 2008-02-01T14:36:35-00:00 2008 Nat Methods 1548-7105 5 2 135 146 expression genomics protein purification structural-genomics structure http://www.citeulike.org/user/neils/article/2221373 <i>Structure, Vol. 16, No. 1. (January 2008), pp. 115-124.</i><br /><br />The activation segment of protein kinases is structurally highly conserved and central to regulation of kinase activation. Here we report an atypical activation segment architecture in human MPSK1 comprising a beta sheet and a large alpha-helical insertion. Sequence comparisons suggested that similar activation segments exist in all members of the MPSK1 family and in MAST kinases. The consequence of this nonclassical activation segment on substrate recognition was studied using peptide library screens that revealed a preferred substrate sequence of X-X-P/V/I-varphi-H/Y-T( *)-N/G-X-X-X (varphi is an aliphatic residue). In addition, we identified the GTPase DRG1 as an MPSK1 interaction partner and specific substrate. The interaction domain in DRG1 was mapped to the N terminus, leading to recruitment and phosphorylation at Thr100 within the GTPase domain. The presented data reveal an atypical kinase structural motif and suggest a role of MPSK1 regulating DRG1, a GTPase involved in regulation of cellular growth. Structure of the Human Protein Kinase MPSK1 Reveals an Atypical Activation Loop Architecture. J Eswaran A Bernad JM Ligos B Guinea JE Debreczeni F Sobott SA Parker R Najmanovich BE Turk S Knapp doi:10.1016/j.str.2007.10.026 Structure, Vol. 16, No. 1. (January 2008), pp. 115-124. 2008-01-12T01:36:36-00:00 2008 Structure 0969-2126 16 1 115 124 activation activity kinase loop mpsk1 specificity structure